Journal: bioRxiv

Article Title: Dual GLP-1 and GIP receptor agonist Tirzapetide plays an off-target role in the modulation of the β-adrenoceptors and glucose metabolism in hyperglycemic or senescent cardiac cells

doi: 10.1101/2025.04.09.648025

Figure Lengend Snippet: (A) The cellular resting level of Ca 2+ was imagined in all groups of H9c2 cells loaded with a Ca 2+ -sensitive fluorescence dye Fluo-3 AM. The representative images demonstrate the effect of TZPD (40 nM) on intracellular Ca 2+ distribution in resting cells with and without receptor antagonism for the SC group of cells (left) and HG group of cells (right). The average cellular Ca 2+ level in Fluo-3 AM loaded H9c2 cells was measured in all groups by the flow cytometer technique. Representative flow cytometry data are given in the left parts. The protein levels of phosphorylated PKA (pPKA) and PKA and their ratios (B), and phosphorylated GSK (pGSK) and GSK and their ratios (C) , compared to a reference protein GAPDH in the SC, SC+TZPD, SC+TZPD+ANT or HG, HG+TZPD, HG+TZPD+ANT comparison to the NC group. All shortens are the same as given in previous figures. Representative protein bands are given in the left parts of the bar graphs. Data are presented as means±SEM. * p <0.05 vs. NC group, γ p <0.05 vs. SC group, δ p <0.05 vs. HG group, Ψ p <0.05 vs. SC+ANT+TZPD group, τ p <0.05 vs. HG+ANT+TZPD group ( n =3-4 independent experiments).

Article Snippet: Following the transfer and blocking steps with 1% BSA in TBS-0.3% Tween, the membrane was incubated with the primer antibodies of either β1-AR (Thermo,), β2-AR (Origine, NM_012492), β3-AR (Saint Jones, STJ91728-200), GLP-1R (MedChem, HY-P81160) GIP-R (MedChem, HY-P10138), PKG (Thermo, PA3-031A), Glut4 (Medchem, HY-P80689), pPKA (Cell signaling, 4781), PKA (Cell signaling, 4782), pGSK (Santa Cruz, sc-373800), GSK (Santa Cruz, sc-377213), eNOS (Thermo, PA3-031A), alpha tubulin (Santa Cruz, sc-5286), and GAPDH (Santa Cruz, sc-137179) primer antibody to use as house-keeping control.

Techniques: Fluorescence, Flow Cytometry, Comparison

Journal: Alzheimer's research & therapy

Article Title: Targeting early tau pathology: probiotic diet enhances cognitive function and reduces inflammation in a preclinical Alzheimer's model.

doi: 10.1186/s13195-025-01674-1

Figure Lengend Snippet: Fig. 3 The effects of htauE14 and probiotic supplement on tau phosphorylation in the LC and levels of associated GSK-3β activation in the hippocampus. A An example image of bilateral AAV infusion sites in the LC. B Co-expression of htauE14 (GFP; green) and dopamine β-hydroxylase (DBH; red) in LC neu rons. Scale bars: 50 μm. C Representative blot of pTau S262 in LC tissue (left) and quantification of relative optical density (ROD; right). D Representative blot of pTau S356 in LC tissue (left) and quantification of relative optical density (ROD; right). N: GFP: 3 M; htauE14: 1 F/1M. E Representative blot of pGSK- 3β across groups in hippocampal tissue. F RODs of pGSK-3β levels. G Comparison of pGSK-3β levels based on diet x sex interaction. H Representative blot of pGSK-3β across groups in hippocampal tissue. I RODs of GSK-3β levels. J Ratios of pGSK-3β/GSK-3β. N: GFP + Ctrl Diet: 4 F/4 M; GFP + P/P Diet: 4 F/4 M; htauE14 + Ctrl Diet: 4 F/4 M; htauE14 + P/P Diet: 4 F/4 M. *p < 0.05, **p < 0.01

Article Snippet: They were then incubated for 2 h at room temperature with the following antibodies: pTau S262 (AB92627, Abcam, 1:2000), pTau S356 (AB75603, Abcam, 1:5000), pGSK-3β (sc-373800, Santa Cruz, 1:1000) and GSK-3β (sc-53831, Santa Cruz, 1:1000).

Techniques: Phospho-proteomics, Activation Assay, Expressing, Comparison

Journal: PLOS ONE

Article Title: Palmitoyl-L-carnitine induces tau phosphorylation and mitochondrial dysfunction in neuronal cells

doi: 10.1371/journal.pone.0313507

Figure Lengend Snippet: (A) shows changes in protein levels of pGSK-3β (S9), GSK-3β, CDK5, and p25 after treatment with palmitoyl-L-carnitine in SH-SY5Y cells. pGSK-3β (S9) indicates GSK-3β phosphorylated at serine 9. Full blots are provided in . (B) Histogram illustrating changes in protein levels of pGSK-3β (S9), CDK5, and p25 after treatment with palmitoyl-L-carnitine in SH-SY5Y cells, shown as mean ± standard error of the mean (SEM; n = 3). GSK-3β phosphorylation levels were normalized to the total GSK-3β protein. (C) shows changes in protein levels of pTau (T181), pTau (S262), PHF-1, total tau, GSK-3β, CDK5, and p25 after treatment with tau kinase inhibitors in palmitoyl-L-carnitine-treated SH-SY5Y cells. pTau (T181), pTau (S262), and PHF-1 represent tau phosphorylated at threonine 181, serine 262, and serine 396/404, respectively. SB216763, Roscovitine, and PD150606 are GSK-3β inhibitor, CDK5 inhibitor, and calpain inhibitor, respectively. Full blots are provided in . (D) Histogram illustrating changes in protein levels of pTau (T181), pTau (S262), PHF-1, total tau, GSK-3β, CDK5, and p25 after treatment with tau kinase inhibitors in palmitoyl-L-carnitine-treated SH-SY5Y cells, shown as mean ± standard error of the mean (SEM; n = 3). Tau phosphorylation levels were normalized to the total tau protein. In (A) and (D) , BSA and BSA-PC represent bovine serum albumin and BSA-conjugated palmitoyl-L-carnitine, respectively. Statistical significance was determined using an unpaired two-tailed t-test with Welch’s correction and an ordinary two-way ANOVA with Tukey’s multiple comparison test; ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The primary antibodies included pTau (T181; CST #12885), pTau (S262; Abcam #ab131354), PHF-1 (Peter Davies Lab.), Tau5 (Peter Davies Lab.), pDRP1 (S616; CST #4494), DRP1 (CST #8570), pMFF (S146; CST #49281), MFF (CST #14739), OPA1 (CST #80471), Mitofusin-1 (CST #14739), Mitofusin-2 (CST #11925), pGSK-3β (S9; CST #5558), GSK-3β (CST #12456), CDK5 (Santa Cruz #sc-173), p35 (Santa Cruz #sc-820), p62 (CST #8025), LC3 (CST #3868), and Actin (Millipore #MAB1501).

Techniques: Phospho-proteomics, Two Tailed Test, Comparison